Evidence that phenol phosphorylation to phenylphosphate is the first step in anaerobic phenol metabolism in a denitrifying Pseudomonas sp.

Lack, Achim; Fuchs, Georg

doi:10.1007/bf00276473

Cited by 46 publications

(57 citation statements)

References 33 publications

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“…In our investigations with the hydroquinone fermenting strain HQG61, CO2 fixation was measurable only as formation of the carboxylated end product benzoate (Gorny and Schink 1994). Carboxylation of phenylphosphate was demonstrated in vitro in cell-free extracts of phenol-degrading nitrate-reducing bacteria (Lack and Fuchs 1994).…”

Section: Discussionmentioning

confidence: 64%

Complete anaerobic oxidation of hydroquinone by Desulfococcus sp. strain Hy5: indications of hydroquinone carboxylation to gentisate

Gorny

Schink

1994

Arch. Microbiol.

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The sulfate-reducing strain Hy5 was able to grow with hydroquinone as sole source of carbon and energy. In experiments with dense cell suspensions, several indications were found that gentisate was the first intermediate in anaerobic degradation of hydroquinone: (1) degradation of hydroquinone was accelerated by addition of bicarbonate; (2) cell suspensions grown with hydroquinone oxidized gentisate at a rate similar to that of suspensions grown with gentisate, whereas the latter were not able to degrade hydroquinone in the presence of chloramphenicol; (3) in SDS-PAGE analysis of cell-free extracts of strain Hy5, two additional protein bands were found after growth with hydroquinone that were not detected in cells grown with gentisate, probably representing a hydroquinone carboxylating enzyme. A corresponding enzyme activity could not be detected. In cell-free extracts of hydroquinone-grown strain Hy5, the specific acyl-CoA ligase activity with gentisate as substrate was detected at 70 nmol • mg -1 x min-L Gentisyl-CoA was enzymatically reduced to several unidentified nonaromatic products in the presence of dithionite-reduced methyl viologen.

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Section: Discussionmentioning

confidence: 64%

Complete anaerobic oxidation of hydroquinone by Desulfococcus sp. strain Hy5: indications of hydroquinone carboxylation to gentisate

Gorny

Schink

1994

Arch. Microbiol.

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“…Phenol-grown cells were harvested by centrifugation (10 500 g; 10 min) and the supernatant was filtered with a 0.2 mm filter. Approximately 30 ml of the supernatant was then loaded onto Whatman flexible plates (250 m layer Al/Sil G/UV), 1 ml at a time, and the intermediates were separated with a solvent consisting of ethanol/dichloromethane/H 2 O (8:1:1) as previously described (Lack and Fuchs, 1994;Schleinitz et al, 2009). The separated compounds were then visualized on an ultraviolet light box.…”

Section: Analytical Techniquesmentioning

confidence: 99%

“…In the case of phenol, two different pathways have been proposed (Figure 3). Several denitrifying species from the genera Pseudomonas, Azoarcus and Thauera, as well as the Fe(III) reducer, Geobacter metallireducens, initially phosphorylate phenol to phenylphosphate, which is then carboxylated to produce 4-hydroxybenzoate (Lack and Fuchs, 1994;Breinig et al, 2000;Rabus et al, 2005;Schleinitz et al, 2009). Clostridium hydroxybenzoicum, on the other hand, directly carboxylates phenol to 4-hydroxybenzoate (Zhang and Wiegel, 1994).…”

Section: Introductionmentioning

confidence: 99%

Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

et al. 2011

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Insight into the mechanisms for the anaerobic metabolism of aromatic compounds by the hyperthermophilic archaeon Ferroglobus placidus is expected to improve understanding of the degradation of aromatics in hot (>80° C) environments and to identify enzymes that might have biotechnological applications. Analysis of the F. placidus genome revealed genes predicted to encode enzymes homologous to those previously identified as having a role in benzoate and phenol metabolism in mesophilic bacteria. Surprisingly, F. placidus lacks genes for an ATP-independent class II benzoyl-CoA (coenzyme A) reductase (BCR) found in all strictly anaerobic bacteria, but has instead genes coding for a bzd-type ATP-consuming class I BCR, similar to those found in facultative bacteria. The lower portion of the benzoate degradation pathway appears to be more similar to that found in the phototroph Rhodopseudomonas palustris, than the pathway reported for all heterotrophic anaerobic benzoate degraders. Many of the genes predicted to be involved in benzoate metabolism were found in one of two gene clusters. Genes for phenol carboxylation proceeding through a phenylphosphate intermediate were identified in a single gene cluster. Analysis of transcript abundance with a whole-genome microarray and quantitative reverse transcriptase polymerase chain reaction demonstrated that most of the genes predicted to be involved in benzoate or phenol metabolism had higher transcript abundance during growth on those substrates vs growth on acetate. These results suggest that the general strategies for benzoate and phenol metabolism are highly conserved between microorganisms living in moderate and hot environments, and that anaerobic metabolism of aromatic compounds might be analyzed in a wide range of environments with similar molecular targets.

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“…The process has been studied in the denitrifying bacterium Thauera aromatica (3,(32)(33)(34)63) (Fig. 1).…”

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Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica

2000

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Genes involved in the anaerobic metabolism of phenol in the denitrifying bacterium Thauera aromatica have been studied. The first two committed steps in this metabolism appear to be phosphorylation of phenol to phenylphosphate by an unknown phosphoryl donor ("phenylphosphate synthase") and subsequent carboxylation of phenylphosphate to 4-hydroxybenzoate under release of phosphate ("phenylphosphate carboxylase"). Both enzyme activities are strictly phenol induced. Two-dimensional gel electrophoresis allowed identification of several phenol-induced proteins. Based on N-terminal and internal amino acid sequences of such proteins, degenerate oligonucleotides were designed to identify the corresponding genes. A chromosomal DNA segment of about 14 kbp was sequenced which contained 10 genes transcribed in the same direction. These are organized in two adjacent gene clusters and include the genes coding for five identified phenol-induced proteins. Comparison with sequences in the databases revealed the following similarities: the gene products of two open reading frames (ORFs) are each similar to either the central part and N-terminal part of phosphoenolpyruvate synthases. We propose that these ORFs are components of the phenylphosphate synthase system. Three ORFs showed similarity to the ubiD gene product, 3-octaprenyl-4-hydroxybenzoate carboxy lyase; UbiD catalyzes the decarboxylation of a 4-hydroxybenzoate analogue in ubiquinone biosynthesis. Another ORF was similar to the ubiX gene product, an isoenzyme of UbiD. We propose that (some of) these four proteins are involved in the carboxylation of phenylphosphate. A 700-bp PCR product derived from one of these ORFs cross-hybridized with DNA from different Thauera and Azoarcus strains, even from those which have not been reported to grow with phenol. One ORF showed similarity to the mutT gene product, and three ORFs showed no strong similarities to sequences in the databases. Upstream of the first gene cluster, an ORF which is transcribed in the opposite direction codes for a protein highly similar to the DmpR regulatory protein of Pseudomonas putida. DmpR controls transcription of the genes of aerobic phenol metabolism, suggesting a similar regulation of anaerobic phenol metabolism by the putative regulator.Phenolic compounds are important plant constituents. For example, lignin represents about 20 to 30% of the biomass of higher plants and consists of phenylpropane units with various hydroxyl or methoxyl groups. Phenol is formed from lignin and several other natural or synthetic substrates during microbial degradation. The best-known phenol-generating enzyme is tyrosine phenol lyase, whose activity is used in bacterial taxonomic studies. Because of its toxic effects on cells, phenol was used as an antiseptic in clinical applications for a long time. In the chemical industry, phenol is a basic substance for the production of phenolic compounds from coal. Because of its broad application in industry, phenolic compounds are among the most prominent man-made groundwater co...

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Evidence that phenol phosphorylation to phenylphosphate is the first step in anaerobic phenol metabolism in a denitrifying Pseudomonas sp.

Cited by 46 publications

References 33 publications

Complete anaerobic oxidation of hydroquinone by Desulfococcus sp. strain Hy5: indications of hydroquinone carboxylation to gentisate

Complete anaerobic oxidation of hydroquinone by Desulfococcus sp. strain Hy5: indications of hydroquinone carboxylation to gentisate

Genome-scale analysis of anaerobic benzoate and phenol metabolism in the hyperthermophilic archaeon Ferroglobus placidus

Genes Involved in Anaerobic Metabolism of Phenol in the Bacterium Thauera aromatica

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